Malaria, tuberculosis, and AIDS are three major pathogen-specific causes of human mortality in the world. More than 90% of the world's malaria is in Africa, where the mosquito Anopheles gambiae is the primary vector. Human and malaria parasite genome projects are now underway and expected to be complete within the next year or two. This proposal seeks to initiate an A.gambiae genome project, with the expectation that the availability of the genomes of the parasite, human host, and the vector will lead to the development of novel malaria control strategies. The immediate aims of this proposal are the following: (1)to sequence 10,000 ESTs from normalized cDNA libraries made from each of five different mosquito tissues (salivary gland, midgut, fatbody, head, and immune- responsive cell lines), (2) to physically map to the mosquito's polytene chromosomes 2,000 genomic DNA Bacterial Artificial Chromosomes (BACs), (3) to sequence both ends of 25,000 genomic DNA BACs, and (4) to assess local scale synteny between the mosquito and Drosophila melanogaster by sequencing 3 megabases of sequence selected to be homologous with the well studied D. melanogaster Adh region. These 4 aims will provide sequences for direct gene discovery, for gene expression analysis (by microarray techniques), and for the determination of broad (polytene chromosome level) and local (megabase sequence level) synteny between D .melanogaster and A. gambiae. They will also facilitate a subsequent full genome sequencing project, either by a Sequence Tagged Connector strategy or by a shotgun strategy. The project is structured to involve the participation of both US and European institutions so as to make this an international effort. The long range is to encourage a broad range of funding agencies to join in supporting an effort to sequence the entire genome of this mosquito.